3473 results about "Target gene" patented technology

A modification template for improving the inhibitory effect of a double-stranded oligonucleotide on a target gene, a combination thereof and the use thereof. The double-stranded oligonucleotide is an siRNA and contains a sense strand and an antisense strand, and each nucleotide of the sense strand and antisense strand is a modified nucleotide. The double-stranded oligonucleotide is modified with a special modification template, and has a significantly improved inhibition rate on a target gene compared with a basic sequence.

The invention belongs to the technical field of RNA biological prevention and treatment, and particularly relates to dsRNA for preventing and treating crop anthracnose and application of the dsRNA. The dsRNA is targeted inositol phosphatidylceramide (IPC) synthetase coding gene dsRNA, and is obtained by transcription of a part of sequence (SEQ ID No.1) of an inositol phosphatidylceramide (IPC) synthetase coding gene CaAUR1, the gene segment is a segment of a target gene for preventing and treating crop anthracnose, and the dsRNA obtained by transcription can obviously reduce pathogenicity of mango anthracnose bacteria, and has the effects of keeping mango fresh, preventing and treating anthracnose. And the compound can be used for preventing and treating the crop anthracnose or preparing products for preventing and treating the crop anthracnose.

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The invention belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody for specific recognition of type II grass carp reovirus VP4 protein, an eukaryotic expression method and application. According to the invention, a target gene S6 is transfected to an HEK293 cell for expression and purification, a New Zealand white rabbit is immunized in cooperation with a Freund's adjuvant, and the rabbit-derived recombinant monoclonal antibody capable of specifically recognizing the GCRV-II VP4 protein is obtained by using ELISA, single B cell screening and eukaryotic expression technologies. The antibody has strong specificity, provides support for further establishment of a specific II-type grass carp reovirus diagnosis technology, and has great application value in development of GCRV-II related scientific research.

The invention provides a plant salt tolerance related transcription factor TaVOZ1 as well as a coding gene and application thereof, and belongs to the technical field of functional genes. The TaVOZ1 is related to plant salt tolerance and has an amino acid sequence as shown in SEQ ID No. 1, and a gene for coding the protein has a nucleotide sequence as shown in SEQ ID No. 2. The plant salt tolerance can be regulated by regulating the content or activity of the protein or the expression quantity of the gene, for example, when the gene is over-expressed, the plant salt tolerance is enhanced, and when the gene is knocked down, the salt tolerance is weakened. The TaVOZ1 disclosed by the invention can be combined with 5 '-CTTCT-3'or a reverse complementary sequence 5'-AAGAAG-3 'in a downstream target gene promoter to activate the expression of a stress response gene so as to improve the salt tolerance of a plant, so that the TaVOZ1 can be used for culturing a salt-tolerant wheat variety.

The present invention relates to T lymphocytes targeting CLD18A2 and a preparation method and application thereof. The inventors have successfully found a target gene CLD18A2 suitable for developing CAR T cells from a number of tumor-related genes for the first time, and successfully prepared CAR T cells targeting CLD18A2, thereby providing a new treatment method for pancreatic cancer, gastric cancer and other tumors.

The invention discloses a modification template for improving the target gene inhibition effect of double-stranded oligonucleotide and a combination and application of the modification template, the double-stranded oligonucleotide is siRNA and contains a positive-sense strand and an antisense strand, and each nucleotide of the positive-sense strand and the antisense strand is a modified nucleotide. The double-stranded oligonucleotide is modified by a special modification template, and compared with a basic sequence, the inhibition rate of the double-stranded oligonucleotide to a target gene is remarkably improved.

The invention provides an intracellular mycobacterium tuberculosis in-situ nucleic acid detection method based on CRISPR / Cas and application, and the method comprises the following steps: S1, designing crRNA and a primer pair of a mycobacterium tuberculosis target gene sequence, and synthesizing a single-stranded DNA fluorescent probe; s2, preparing a standard solution containing a target gene sequence, and incubating with a CRISPR / Cas12a system; s3, extracting nucleic acids of different types of bacteria, uniformly mixing the nucleic acids of different types of bacteria with the CRISPR / Cas12a system, and incubating; s4, incubating an attenuated strain of mycobacterium tuberculosis and the DNA fluorescent probe, infecting macrophages, and incubating to obtain infected cells; s5, electrically transferring a CRISPR / Cas12a system into the infected cells in S4 by utilizing an electroporation technology, incubating, and detecting the intensity of a fluorescence signal; and S6, performing quantitative and positioning analysis on the detected fluorescence signal to complete detection, and directly introducing a CRISPR / Cas12a system into the macrophage by adopting an electroporation technology to realize intracellular in-situ detection, thereby avoiding the operation complexity of a nucleic acid extraction link and the loss of detection sensitivity in the prior art.

The invention provides a cell specific transcription factor regulatory network analysis method and a visualization platform, and relates to the technical field of bioinformatics, the method comprises the following steps: constructing a gene regulatory network through a GRNBoost2-cisTarget-AUCell-Cell GRN workflow based on a transcription factor in combination with a motif database; screening a direct regulation relationship in combination with the database, and calculating an activity score of a regulator in each cell; based on activity scores and cell type annotation results, grouping the single cell data by using a unified manifold approximation and projection (UMAP) dimensionality reduction method and a Leiden clustering algorithm, and displaying the following results through an interactive visualization tool: a cell clustering UMAP graph, performing color marking according to cell types; a UMAP graph and a heat map of transcription factor regulator activity; according to the visual map of the gene regulation and control network, transcription factors and target genes are distinguished through node shapes, and regulation and control relations are marked through line weights and colors. According to the invention, an accurate regulation and control network can be provided.

The invention discloses a soybean bidirectional promoter and application thereof, and belongs to the technical field of separation and application of bidirectional promoters. The nucleotide sequence of the soybean bidirectional promoter disclosed by the invention is as shown in SEQ ID NO. 2. Experiments prove that the soybean bidirectional promoter can bidirectionally and simultaneously drive the expression of a target gene, and by applying the soybean bidirectional promoter, plant traits can be improved, transgenic plants or new plant varieties can be cultivated, and the biological breeding process can be promoted.

The invention relates to a gene editing vector of agrobacterium, a gene editing method of the gene editing vector and an agrobacterium editing strain. The agrobacterium gene editing vector comprises the following gene elements: a plasmid skeleton, a Pvan promoter, a spCas9 gene, a green fluorescent protein gene GFP, gRNA, and an upstream homologous arm and a downstream homologous arm of a target gene. According to the agrobacterium gene editing vector, a CRISPR / spCas9-GFP gene editing system is adopted, an efficient, simple, convenient and traceless genome editing system is constructed in agrobacterium, and gene knockout and insertion can be achieved.

The invention discloses a double-stranded RNA (Ribonucleic Acid) aiming at hemiptera insects as well as a pesticide composition and application of the double-stranded RNA. The dsRNA provided by the invention is used as a biopesticide, has the characteristics of high efficiency, specificity, fast degradation and no residue, realizes the prevention and control of brown planthopper through an RNAi mechanism, and is beneficial to reducing the use of chemical pesticides. The invention provides a brand new brown planthopper target gene as a target of RNA interference. The survival rate result shows that the genes play an important role in the survival process of the brown planthopper, are effective targets for preventing and treating the brown planthopper by using a gene silencing means, and have a wide application prospect. Correspondingly, the invention provides a plurality of dsRNA sequences aiming at the targets, and part of the sequences show unprecedented brown planthopper control efficiency.

The invention relates to the technical field of plant genetic engineering, in particular to a promoter and application thereof in orchid plants. The invention provides an orchid endogenous promoter which is a powerful constitutive promoter and can drive a target gene to be powerfully expressed in various tissues such as roots, stems, leaves and flowers of an orchid. Compared with a CaMV 35S promoter, the promoter has the advantages that the driving efficiency of the gene in orchid plants is greatly improved, an important tool is provided for molecular breeding of plants, and the promoter has a relatively good application prospect.

The invention discloses a tomato LACS2 gene promoter and application thereof, and belongs to the technical field of biology. On one hand, the invention provides the tomato LACS2 gene promoter, and on the other hand, the invention provides application of the tomato LACS2 gene promoter in promoting target gene expression in plants. According to the invention, a tomato LACS2 gene promoter fragment (788bp) is constructed to drive an EGFP-GUS (Enhanced Green Fluorescent Protein-Glucuronidase) gene fusion expression vector. GUS histochemical staining and laser confocal microscopy observation prove that the promoter fragment can guide normal expression of the reporter gene in the tomato epidermis. Therefore, the LACS2 promoter can be used for researching development, quality improvement and postharvest preservation of tomato peels and specific expression of exogenous genes in the tomato peels.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a macrophage Angulin-1 gene and / or an Angulin-1 protein in preparation of medicines and diagnostic products for preventing and / or treating atherosclerosis and related diseases. Research finds that plaque and necrotic core areas of an atherosclerosis animal model are remarkably increased due to macrophage Angulin-1 gene knockout, and disease development is promoted; and the recovery of the expression level of Angulin-1 significantly inhibits the development of lesion. Cell experiments show that Angulin-1 reduces intake of oxidized low-density lipoprotein by down-regulating expression of macrophage LOX-1, so that formation of foam cells is inhibited. Therefore, the macrophage Angulin-1 can be used as an atherosclerosis drug target, a gene therapy target gene and an auxiliary diagnosis marker, and a new theoretical basis and an intervention strategy are provided for prevention and treatment of the disease.

The present invention belongs to the field of biotechnology, and specifically relates to a GmSGT2 gene related to plant height and branch development, and a mutant thereof and the use thereof. A Glycine max mutant having fewer branches and decreased plant height is obtained by means of EMS mutagenesis, and a target gene thereof that is located by means of a map-based cloning technique is a GmSGT2 gene of Hedou 12. It is found through searching that the gene encodes soyasapogenol B glucuronide galactosyltransferase, and can galactosylate soyasapogenol B monoglucuronide, thereby affecting the plant height of Glycine max and reducing the branches of Glycine max. Therefore, the GmSGT2 mutant can be used for cultivating dwarf high-yield Glycine max varieties, and is of great significance in the breeding of ideal plant types of Glycine max and research on important agronomic traits of plants. The mutant has broad application prospects and great research value with regard to understanding the plant height and branch regulation mechanism of Glycine max and improving the process of Glycine max breeding; and can provide an excellent germplasm resource stock for regulating the close planting and high-yield breeding of Glycine max.

The embodiment of the invention provides a method for regulating and controlling the form of ginseng adventitious roots and saponin accumulation. The method comprises the following steps: (a) carrying out mutagenesis treatment on initial ginseng adventitious roots by using ethyl methane sulfonate (EMS); (b) performing multiple rounds of screening and subculture to obtain to-be-selected ginseng adventitious roots with different forms and saponin accumulation; and (c) determining one or more target genes for regulating and controlling the form of the ginseng adventitious root and saponin accumulation based on the ginseng adventitious root to be selected.

The invention provides a branched chain amino acid transaminase gene and application thereof, and belongs to the field of gene engineering. According to the invention, the branched chain amino acid transaminase gene HvBCAT5 is found for the first time, and the gene has obvious positive correlation with the accumulation of valine (Val), isoleucine (Ile) and leucine (Leu) of crops. The invention provides an important target gene HvBCAT5 for crop quality improvement, and has important value for increasing the content of branched chain amino acid of crops and improving dietary nutrition. The HvBCAT5 gene as well as a recombinant vector, a recombinant bacterium and a transgenic plant thereof have good application prospects in the fields of food and feed industry.

The invention discloses a phoebe zhennan PzGGPS12 gene and application of the phoebe zhennan PzGGPS12 gene to improvement of yield and drought tolerance of plant terpenoids, and belongs to the field of genetic engineering, a key enzyme gene PzGGPS12 for catalyzing biosynthesis of the terpenoids in phoebe zhennan wood is screened and analyzed, and a full-length CDS nucleic acid sequence of the PzGGPS12 is obtained. Transgenic experiments prove that overexpression of the gene improves the relative content of tobacco monoterpenes and triterpenes. Short-term drought experiments prove that overexpression of the gene can promote biosynthesis of sesquiterpenes, diterpenes and triterpenes so as to enhance drought tolerance of transgenic tobacco. Therefore, the gene can be introduced into a plant as a target gene, the yield of terpenoids in the plant is increased, and the drought tolerance of the plant is enhanced so as to improve the variety of the plant.

The invention relates to cloning of a stress-resistant gene LpbHLH144 of Lilium pumilum DC (Lilium pumilum DC.) and application of the stress-resistant gene LpbHLH144 of Lilium pumilum DC. Comprising the following steps: (1) cloning a target gene; (2) constructing an expression vector; and (3) detecting physiological indexes of the transgenic plant. According to the experiment, lilium tenuifolium is taken as a research object, and the LpbHLH144 gene is cloned by applying an RT-PCR technology; transforming tobacco by constructing an overexpression vector; the functions of overexpression of the LpbHLH144 for improving the saline-alkaline and drought stress of the tobacco are identified through transgenic tobacco, the functions of the lilium tenuifolium bHLH gene when the lilium tenuifolium bHLH gene is subjected to adversity stress are disclosed, a foundation is laid for further application of the lilium tenuifolium in lily breeding, and a theoretical basis is provided for the functions and effects of bHLH transcription factors.

The technology described herein relates to 5'-modified nucleosides, nucleotides, oligonucleotides and double-stranded RNAs, e.g., siRNAs, and kits comprising them and methods of their use for inhibiting target genes.

The invention belongs to the technical field of biology, and particularly relates to an SLC44A1 gene and / or SLC44A5 gene knockout animal model as well as a construction method and application thereof. The construction method of the animal model comprises the following steps: deleting exon regions from the fourth exon region to the thirteenth exon region of the SLC44A1 gene and / or the second exon region of the SLC44A5 gene in mouse brain oligodendroglia cells in a high-efficiency manner by utilizing gene editing and a Cre-loxP recombinase system, so as to realize the knockout of a target gene. The animal model constructed by the invention can be used for researching the regulation mechanism of the choline transporters (SLC44A1 and SLC44A5) on myelin sheath formation, deeply knowing the biological functions of the choline transporters (SLC44A1 and SLC44A5), simulating the occurrence and development processes of human related diseases, revealing the pathogenesis of the related diseases, providing a new thought for prevention and treatment of brain myelin sheath dysplasia, and providing a new foundation for the prevention and treatment of brain myelin sheath dysplasia. And an effective drug or a treatment mode aiming at the disease is screened by utilizing a gene knockout animal model, so that a new choice is provided for clinical treatment of the disease.

The invention relates to the technical field of gene engineering. The invention provides a functional sequence shRNA1 (short hairpin ribonucleic acid 1) for silencing a circular RNAh-hsacirc005838, and further provides a lentiviral vector of the functional sequence shRNA1 and a construction method of the lentiviral vector. The invention also provides a lentivirus packaging titer kit containing the lentiviral vector with the functional sequence shRNA1, and also provides an application of the lentiviral vector with the functional sequence shRNA1 or the lentiviral vector with the functional sequence shRNA1 in preparation of a kit for breast cancer diagnosis. According to the invention, an interference target sequence siRNA1 is obtained by aiming at the annular target gene RNA h-hsacirc005838, then the finally obtained functional sequence shRNA1 can directly and efficiently infect cells and silence the annular RNAh-hsacirc005838 sequence, and the transfection effect is stable.

The invention belongs to the technical field of genetic engineering, and particularly relates to a CsECT protein related to plant disease resistance, a recombinant vector and application of the CsECT protein. The invention provides a CsECT protein related to plant disease resistance, the CsECT protein is synthesized by gene CsECT coding, it is speculated that the gene CsECT is used as an m6A reader, metabolism of target mRNA of the gene CsECT is affected, the stability of disease resistance related target genes is improved, expression of disease resistance genes is promoted, and then the disease resistance of plants is improved. The result of the embodiment shows that after the CsECT gene is cloned, the CsECT transgenic citrus is constructed, the CsECT gene is excessively expressed by the transgenic citrus, the canker resistance of the citrus can be remarkably improved, and then the disease resistance of the citrus is improved. Therefore, the CsECT protein synthesized by the gene CsECT disclosed by the invention can be used for improving the disease resistance of plants.

The invention relates to methods for culturing cells, and cultured animal cells for use in described method. Also provided is a modified animal cell and guide RNAs targeting genes to be modified in the modified and cultured cells of the invention.

The invention discloses an oilseed rape BnaA09.SCL28 gene and a promoter and application thereof. The sequence of the oilseed rape BnaA09.SCL28 gene is shown as SEQ ID No.6, and the sequence of the promoter of the oilseed rape BnaA09.SCL28 gene is shown as SEQ ID No.5. Overexpression of the oilseed rape BnaA09.SCL28 gene can increase the size of oilseed rape seeds, increase the seed yield of an oilseed rape single plant, increase the number of effective siliques of oilseed rape, increase the number of seeds per silique of oilseed rape, and increase the height and branching number of oilseed rape plants. The BnaA09.SCL28 gene promoter can drive a target gene to be specifically expressed in buds, siliques or seeds in the early stage of a plant, which indicates that BnaA09.SCL28 can positively regulate longitudinal growth, branch development and reproductive organ formation of the plant, and provides theoretical support for increasing the yield of a single rape plant.

The invention provides a method for simultaneously detecting three hematogenous parasites based on RAA-LFS and application of the method. The method comprises the following steps: firstly, screening target genes of genomic DNA of specific amplified toxoplasma gondii, babesia and plasmodium and primers of the target genes; a primer and probe combination for specifically amplifying the 18s rRNA-1 gene of the toxoplasma gondii, the 18s rRNA-2 gene of the babesia and the 18s rRNA-3 gene of the plasmodium is determined; rapid multiple detection is completed in a reaction system by utilizing an RAA constant-temperature amplification technology, and a detection result is intuitively judged by combining a lateral flow chromatography test strip, so that the detection efficiency is greatly improved, and the result can be obtained in a short time. The RAA-LFS detection method provided by the invention has remarkable advantages in application scenes such as epidemic disease prevention and control or blood screening.

The invention relates to the field of gene detection, and discloses a detection primer for an abdominal aortic aneurysm related SNP site rs116309149 and application of the detection primer, and the detection primer comprises two non-specific outer side primers and two inner side primers for specifically detecting SNP mutation sites; the outer primer has sequences as shown in SEQ ID NO: 1 and SEQ ID NO: 2 or complementary sequences of the sequences as shown in SEQ ID NO: 1 and SEQ ID NO: 2; and the inner side primer has sequences as shown in SEQ ID NO: 3 and SEQ ID NO: 4 or complementary sequences of the sequences as shown in SEQ ID NO: 3 and SEQ ID NO: 4. According to the detection primer and the application design of the detection primer, the primer which is specifically matched with the rs116309149 site based on an ARMS-PCR method is verified, a reaction system and reaction conditions are perfected and optimized based on the detection primer, amplification and detection of a target gene are successfully completed, and the positive significance is achieved for clinic.

The invention provides an hpRNA interference vector for laver and application of the hpRNA interference vector, and belongs to the technical field of genetic engineering and algae biology. According to the hpRNA interference vector, a binary vector pBI121 is used as a skeleton vector, a laver endogenous Actin promoter and a multiple cloning site segment are inserted, the multiple cloning sites allow target gene segments to be cloned in a forward and reverse repetition mode, the target gene segments are spaced by introns, and therefore an efficient hpRNA structure is formed after transcription, and the efficient hpRNA interference vector is obtained. The gene can be directly used for silencing important genes related to growth and development, metabolic regulation, stress response and the like in the laver. The invention further provides a construction method of the hpRNA interference vector and application of the hpRNA interference vector in silencing laver genes and provides a method for silencing laver gene expression, the gene silencing effect is remarkable, experimental results can be repeated, and a powerful tool is provided for molecular breeding and genetic improvement of laver.

The invention discloses a carbapenem drug resistance gene detection reagent or kit, the detection reagent comprises a PCR detection liquid, the PCR detection liquid comprises upstream and downstream primer sequences and probe sequences of target genes, and the target genes are KPC, OXA-23, NDM, VIM and IMP. The invention also discloses application of the reagent or the kit. The reagent / kit provided by the invention has simple components, only comprises a PCR detection liquid and negative and positive quality control, can obtain detection results of five major carbapenem drug resistance genes, namely KPC, OXA-23, NDM, VIM and IMP, within about 50 minutes only once, and is simple and convenient to operate, low in detection cost, short in detection time and high in sensitivity. In addition, the lowest detectable concentration of detection of carbapenem drug resistance genes KPC, OXA-23, NDM, VIM and IMP is 10 copy / mL, and the sensitivity is high.